Structure of SARS Coronavirus Spike Glycoprotein

SARS 冠状病毒刺突糖蛋白的结构

• 批准号: 6758154
• 负责人: MICHAEL JEFFREY ROOT
• 金额: $ 23.4万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6758154
• 关键词: SARS virus; X ray crystallography; antiviral agents; biophysics; cell line; conformation; drug discovery /isolation; emerging infectious disease; glycoproteins; host organism interaction; mass spectrometry; peptide chemical synthesis; protein structure; protein structure function; proteolysis; recombinant proteins; severe acute respiratory syndrome; structural biology; transfection; virus infection mechanism; virus protein

DESCRIPTION (provided by applicant): Severe acute respiratory syndrome (SARS) has recently emerged as a new human disease with a mortality rate as high as 15%. The genome of the putative etiological agent, SARS-associated coronavirus (SCV), contains a spike glycoprotein that is implicated in viral attachment and entry by homology to similar proteins in related coronaviruses. These spike glycoproteins contain two regions of hydrophobic heptad repeat sequences characteristic of the viral trimer-of-hairpins structural motif. Formation of a trimer-of-hairpins is an essential step in the membrane fusion process of other enveloped viruses, including HIV-1, Ebola virus and respiratory syncytial virus. Moreover, for these unrelated viruses, polypeptides and small molecules that disrupt the formation of this crucial structure are potent inhibitors of viral infectivity. The studies proposed here test the trimer-of-hairpins structural hypothesis for SCV and develop membrane fusion assays to test possible inhibitors of SCV spike glycoprotein. The specific aims are: 1) To generate and biophysically characterize polypeptides derived from the heptad repeat regions of SCV spike glycoprotein, and to solve the structure of particularly stable constructs using x-ray crystallography; and 2) To develop a cell-cell fusion assay and/or a pseudoviral infectivity assay to assess the fusion activity of the SCV spike glycoprotein, and to test polypeptides derived from the heptad repeat regions for potential inhibitory activity. These experiments will explore the structure and function of the SCV spike glycoprotein, essential information for the future development of antiviral therapeutics and vaccines.

描述（由申请人提供）：严重的急性呼吸综合征（SARS）最近成为一种新的人类疾病，死亡率高达15％。推定病因学剂，与SARS相关的冠状病毒（SCV）的基因组含有一种尖峰糖蛋白，该糖蛋白与病毒附着和同源性与相关冠状病毒中的类似蛋白的进入有关。这些尖峰糖蛋白包含两个疏水性七个区域的重复序列，特征是病毒三聚体结构基序。发射剂的形成是其他包膜病毒的膜融合过程的重要步骤，包括HIV-1，埃博拉病毒和呼吸道合成病毒。此外，对于这些无关的病毒，多肽和小分子破坏了这种关键结构的形成是病毒感染性的有效抑制剂。这里提出的研究测试了SCV的发射剂结构假设，并开发膜融合测定法，以测试SCV SPIKE糖蛋白的可能抑制剂。具体目的是：1）生成和生物物理表征从SCV尖峰糖蛋白的Heptad重复区域得出的多肽，并使用X射线晶体学求解特别稳定的构建体的结构； 2）开发细胞细胞融合测定和/或假病毒感染性测定法，以评估SCV尖峰糖蛋白的融合活性，并测试源自七重复区域的多肽，以获得潜在的抑制活性。这些实验将探讨SCV峰值糖蛋白的结构和功能，这是抗病毒疗法和疫苗未来开发的基本信息。